Generation and functional characterization of anti-human and anti-mouse IL-36R antagonist monoclonal antibodies

Abstract

Deficiency of interleukin (IL)-36 receptor antagonist (DITRA) syndrome is a rare autosomal recessive disease caused by mutations in IL36RN. IL-36R is a cell surface receptor and a member of the IL1R family that is involved in inflammatory responses triggered in skin and other epithelial tissues. Accumulating evidence suggests that IL-36R signaling may play a role in the pathogenesis of psoriasis. Therapeutic intervention of IL-36R signaling offers an innovative treatment paradigm for targeting epithelial cell-mediated inflammatory diseases such as the life-threatening psoriasis variant called generalized pustular psoriasis (GPP). We report the discovery and characterization of MAB92, a potent, high affinity anti-human IL-36 receptor antagonistic antibody that blocks human IL-36 ligand (α, β and γ)-mediated signaling. In vitro treatment with MAB92 directly inhibits human IL-36R-mediated signaling and inflammatory cytokine production in primary human keratinocytes and dermal fibroblasts. MAB92 shows exquisite species specificity toward human IL-36R and does not cross react to murine IL-36R. To enable in vivo pharmacology studies, we developed a mouse cross-reactive antibody, MAB04, which exhibits overlapping binding and pharmacological activity as MAB92. Epitope mapping indicates that MAB92 and MAB04 bind primarily to domain-2 of the human and mouse IL-36R proteins, respectively. Treatment with MAB04 abrogates imiquimod and IL-36-mediated skin inflammation in the mouse, further supporting an important role for IL-36R signaling in epithelial cell-mediated inflammation.

IL‐36 receptor antagonistic antibodies inhibit inflammatory responses in preclinical models of psoriasiform dermatitis

“…These data are consistent with results from recent studies in which anti‐IL‐36R antibodies ameliorated imiquimod‐induced psoriasiform dermatitis in mice and reduced the inflammatory responses from human psoriatic lesional skin transplanted onto immunodeficient mice to a degree similar to etanercept . The differences in the efficacy observed in the imiquimod model and the IL‐23 model could possibly be due to the fact that IL‐36 drives effects at the level of the keratinocyte which is upstream of the IL‐23 model which directly affects T cells . Collectively, these data, including the overt dermatitis induced by the dermal injection of IL‐36α, support the hypothesis that the IL‐36 signalling pathway is a contributing pathogenic factor in psoriasis based on their pathway enrichment analysis…”

“…Overall, these data suggest that IL‐36 cytokines contribute to the pathophysiology of IL‐23‐induced psoriasiform dermatitis. These data are consistent with results from recent studies in which anti‐IL‐36R antibodies ameliorated imiquimod‐induced psoriasiform dermatitis in mice and reduced the inflammatory responses from human psoriatic lesional skin transplanted onto immunodeficient mice to a degree similar to etanercept . The differences in the efficacy observed in the imiquimod model and the IL‐23 model could possibly be due to the fact that IL‐36 drives effects at the level of the keratinocyte which is upstream of the IL‐23 model which directly affects T cells .…”

“…Direct intradermal injection of unprocessed IL‐36α (5 μg of rmIL‐36α every other day for 10 days) into dorsal skin of CD1 mice results in chemokine/cytokine expression, leucocyte infiltration (mainly myeloid cells) and acanthosis . Recently, it was reported that direct intradermal injection of IL‐36 cytokines (combination of α, β and γ) into mouse ear skin produced ear swelling as observed in the current study. The major component of inflammatory infiltration from our observations was also myeloid cells, specifically macrophages, which is similar to that in the IL‐23 model .…”

Monogenic autoinflammatory disorders: beyond the periodic fever

“…63 Two rare dermatological conditions, multiple selfhealing palmoplantar carcinoma and familial keratosis lichenoides chronica, have recently been linked to activating heterozygous mutations in NLRP1. 82 The clinical response to targeting IL-1β or NLRP1 was not examined in this small cohort.…”

“…[71][72][73][74][75][76][77][78][79][80][81] Therapeutic targeting of the IL-36 receptor specifically is currently being investigated. 82…”